Ets2 and Mesp1 generate cardiac progenitors from fibroblasts
A method for modulating cell differentiation capabilities using heterologous gene expression. Some embodiments of the invention relate to a method for inducing a cardiac progenitor cell by delivering a reprogramming factor to the cell, wherein the reprogramming factor comprises ETS2 or a combination of ETS2 and Mesp1.
1. An ETS2 treated cell obtained by a method comprising the step of transducing a somatic cell with ETS2 to produce an artificially increased first level of an ETS2 protein in the somatic cell; wherein the artificially increased first level of ETS2 protein transforms the somatic cell into the ETS2 treated cell having surface marker protein levels of SSEA-3 or Tra-1-81 higher than those found in the somatic cell, wherein the somatic cell is a normal human dermal fibroblast cell.
2. The ETS2 treated cell of claim 1 , wherein the ETS2 protein comprises the human ETS2 protein sequence of SEQ ID NO:8 encoded on a DNA expression vector transduced in the somatic cell, wherein the human ETS2 protein sequence is expressed in the somatic cell, whereby the first level of the ETS2 protein is artificially increased.
3. The ETS2 treated cell of claim 2 , wherein the DNA expression vector comprises the ETS2 DNA coding sequence of SEQ ID NO:9.
4. The ETS2 treated cell of claim 1 , wherein an artificially increased first level of an ETS2 protein in the somatic cell transforms the somatic cell into the ETS2 treated cell expressing pluripotent marker genes consisting of NANOG, OCT3/4, SOX2 or c-MYC.
5. The ETS2 treated cell of claim 2 , wherein the DNA expression vector comprises a lentiviral vector introduced by infection.
6. A cardiac progenitor cell obtained by a method comprising the step of transducing the ETS2 treated cell of claim 1 with Mesp1 to produce an artificially increased first level of an Mesp1 protein; wherein the artificially increased first level of ETS2 protein and the artificially increased first level of Mesp1 protein transforms the ETS2 treated cell into the cardiac progenitor cell having levels of contractile protein gene expression of alpha MHC and troponin T higher than those found in the ETS2 treated cell.
7. The cardiac progenitor cell of claim 6 , wherein the Mesp1 protein comprises the mouse Mesp1 protein sequence of SEQ ID NO:5 encoded on a DNA expression vector transduced in the ETS2 treated cells, wherein the mouse Mesp1 protein sequence is expressed in the ETS2 treated cells, whereby the first level of the Mesp1 protein is artificially increased.
8. The cardiac progenitor cell of claim 7 , wherein the DNA expression vector comprises the Mesp1 DNA coding sequence of SEQ ID NO:6.
9. The cardiac progenitor cell of claim 7 , wherein the DNA expression vector comprises a lentiviral vector introduced into the ETS2 treated cells by infection.
10. An induced cardiac progenitor cell obtained by a method comprising the steps of:
(a) transducing a somatic cell with an ETS2 reprogramming factor forming an ETS2 treated cell, wherein the ETS2 treated cell produces a pluripotent stem cell marker protein not present in the somatic cell; and
(b) transducing the ETS2 treated cell with a Mesp1 reprogramming factor forming a cardiac progenitor cell, wherein the cardiac progenitor cell produces a cardiac cell marker protein neither expressed in the somatic cell nor in the ETS2 treated cell;
wherein the somatic cell is a normal human dermal fibroblast cell.